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目的 探讨反义热休克蛋白 70 (hsc70 )在X射线照射下对野生型p5 3(wtp5 3)的影响。方法 利用脂质体方法将本室构建的反义hsc70真核表达重组质粒pXAhsc70转染wtp5 3的MCF 7乳腺癌细胞 ,PCR证实外源DNA存在。Westernblot检测hsc70蛋白下降程度。X射线照射转染细胞后 ,检测wtp5 3蛋白的表达情况及半衰期变化。结果 转染重组质粒的细胞中hsc70蛋白下降 5 3% ;不同剂量X射线照射下 ,反义hsc70的存在能够提高wtp5 3的表达 ;4Gy相同剂量照射后 ,转染反义hsc70重组质粒的细胞中wtp5 3的半衰期延长。结论 反义hsc70能够上调wtp5 3的表达、稳定X射线照射下的wtp5 3蛋白并延长其半衰期 ;推测hsc70可能参与了wtp5 3的稳定和降解过程
Objective To investigate the effect of antisense heat shock protein 70 (hsc70) on wild-type p53 (wtp53) under X-irradiation. Methods The antisense hsc70 eukaryotic expression recombinant plasmid pXAhsc70 constructed in our laboratory was transfected into wtp5 3 MCF 7 breast cancer cells by liposome method, and the presence of exogenous DNA was confirmed by PCR. Western blots detected the degree of hsc70 protein decline. After transfection of cells with X-irradiation, the expression of wtp5 3 protein and changes in half-life were detected. Results The hsc70 protein was decreased by 53% in the cells transfected with the recombinant plasmid; the antisense hsc70 could increase the expression of wtp53 in cells irradiated with different doses of X-rays; after irradiation with the same dose of 4Gy, the cells transfected with antisense hsc70 recombinant plasmid were transfected into the cells. The half-life of wtp5 3 is prolonged. Conclusion Antisense hsc70 can up-regulate the expression of wtp5 3, stabilize the wtp5 3 protein under X-ray irradiation and prolong its half-life. It is speculated that hsc70 may be involved in the stabilization and degradation of wtp5 3.